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Updated: May 12, 2026
Isolation of Primary Mouse Retinal Pigmented Epithelium Cells
Published on: November 4, 2022
In vitro evidence for mycophenolic acid dose-related cytotoxicity in human retinal cells
Leandro C Zacharias1, Francisco Max Damico, Maria C Kenney
1*Department of Ophthalmology, University of São Paulo Medical School, São Paulo, Brazil; †Department of Ophthalmology, School of Medicine, University of California, Irvine, California; ‡Department of Psychology, Psychology Institute, University of São Paulo, São Paulo, Brazil.
Purpose:
Mycophenolic acid (MPA) is an immunosuppressive agent that controls noninfectious uveitis. Intravitreal MPA delivery may be a potential adjuvant therapy in patients who have to discontinue steroid or immunosuppressive systemic therapy because of side effects. The aims of this study are to evaluate the in vitro effects of MPA over human retinal pigment epithelium (ARPE-19) and human Muller cells (MIO M-1).
Methods:
ARPE-19 cells and MIO M-1 cells were exposed to 25, 50, and 100 µg/mL of MPA (Roche Bioscience, Palo Alto, CA) for 24 hours. Toxicity was evaluated by trypan blue dye-exclusion cell viability assay, caspase-3/7 apoptosis-related assay, and JC-1 mitochondrial membrane potential assay.
Results:
The MPA (25 µg/mL and 50 µg/mL) did not cause reduction in cell viability or significant change in caspase-3/7 activity in both cell lines tested. Mycophenolic acid (100 µg/mL) caused a significant decrease in cell viability (P < 0.01) and higher caspase-3/7 activity (P < 0.05) in both cell lines compared with untreated cells. The JC-1 mitochondrial membrane potential did not show statistically significant differences for both cell lines and all concentration tested when compared with untreated controls (P > 0.05).
Conclusion:
Intraocular delivery may be a potential alternative for the treatment of noninfectious uveitis, either by intravitreal injection or sustained-release drug-delivery systems, in doses of 50 µg/mL or lower.
Insights
Mycophenolic acid (MPA) is safe for retinal cells at doses up to 50 µg/mL, suggesting potential for intraocular delivery in treating noninfectious uveitis. Higher doses of MPA showed toxicity in retinal cell lines.
Area of Science:
- Ophthalmology
- Immunology
- Cell Biology
Background:
- Noninfectious uveitis is managed with systemic immunosuppressants.
- Systemic therapies can cause adverse effects, necessitating alternative treatments.
- Intravitreal drug delivery offers localized treatment with potentially fewer side effects.
Purpose of the Study:
- To evaluate the in vitro effects of mycophenolic acid (MPA) on human retinal pigment epithelium (ARPE-19) and Muller cells (MIO-M1).
- To assess the potential of intravitreal MPA as an adjuvant therapy for noninfectious uveitis.
Main Methods:
- ARPE-19 and MIO-M1 cells were exposed to MPA at concentrations of 25, 50, and 100 µg/mL for 24 hours.
- Cell viability was assessed using trypan blue dye exclusion.
- Apoptosis was evaluated via caspase-3/7 assay, and mitochondrial membrane potential using the JC-1 assay.
Main Results:
- MPA at 25 and 50 µg/mL did not significantly affect cell viability or apoptosis.
- MPA at 100 µg/mL significantly decreased cell viability and increased caspase-3/7 activity in both cell lines (P < 0.01 and P < 0.05, respectively).
- Mitochondrial membrane potential remained unaffected across all tested MPA concentrations.
Conclusions:
- Intraocular delivery of MPA at doses of 50 µg/mL or lower is potentially safe for retinal cells.
- This suggests intravitreal MPA may be a viable treatment option for noninfectious uveitis.
- Further research into sustained-release systems for intraocular MPA delivery is warranted.
